Recently I bought MitoTracker Deep Red and PicoGreen from MCE. I prepared MitoTracker Deep Red with 92 μL of DMSO to prepare a 1 mM stock solution, and stored it according to the COA conditions provided on the MCE website. The MitoTracker Deep Red stock solution was diluted to the working solution at 1:6000 with serum-free medium.
Then I prepared the shrimp cells, a total of 6 samples. After cultured for 24 hours, the cells were first incubated with 1 mL of 1X PicoGreen for 40 min, washed 2-3 times with serum-free medium, and then incubated with 1 mL of MitoTracker Deep Red working solution for 30 min. However, it was found that the cells could not be stained with two dyes at the same time. What is the reason? Any suggestions would be greatly appreciated.
Marine Ma Hi, We evaluated two dyes used in your experiments.
PicoGreen emits fluorescence only upon binding to double-stranded DNA;
however, it has limited cell membrane permeability, resulting in stronger fluorescence signals in dead cells (DOI: 10.4236/jst.2016.63003).
In contrast, MitoTracker Deep Red is suitable for live cell staining, which may lead to their fluorescence signals appearing in different cell populations.
Additionally, product information from other biotech suppliers also advises against using PicoGreen for live cell staining and recommends its application primarily for DNA quantification assays.
The answer to this question comes from MedChemExpress (MCE) Technical Support. [email protected]
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